Targeted insertion of exogenous DNA into the eukaryotic genome by the Cre recombinase.
Cre is a 38-kD protein from bacteriophage P1 that catalyzes site-specific recombination between 34-bp loxP sequences. Our previous work has shown that Cre can perform site-specific excisive recombination not only in prokaryotes, but also in eukaryotes such as yeast and cultured mammalian cells. In this work we show that intermolecular Cre-mediated recombination can specifically direct the integration of a loxP-containing circular DNA into a chromosomal loxP site, both in yeast and in mammalian cells. The resulting integrants are predominantly simple single-copy insertions. Cre-mediated recombination thus provides a simple way to direct single-copy site-specific integration of exogenous DNA into the eukaryotic genome.
Sauer
B
B
E.I. DuPont de Nemours & Co., Inc., Central Research and Development Department, Wilmington, DE 19880-0328.
Henderson
N
N
D000818:Animals; D001435:Bacteriophages / Q000201:enzymology; D001483:Base Sequence; D002460:Cell Line; D002875:Chromosomes / Q000648:ultrastructure; D004247:DNA / Q000378:metabolism*; D004254:DNA Nucleotidyltransferases / Q000235:genetics*; D005057:Eukaryotic Cells / Q000378:metabolism; D019426:Integrases; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids; D011995:Recombination, Genetic; D012045:Regulatory Sequences, Nucleic Acid; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013937:Thymidine Kinase / Q000096:biosynthesis / Q000235:genetics; D014170:Transformation, Genetic; D014764:Viral Proteins
D014764:Viral Proteins; D004247:DNA; D013937:Thymidine Kinase; C045073:Cre recombinase; D004254:DNA Nucleotidyltransferases; D019426:Integrases